LOCUS       Exported                6176 bp ds-DNA     circular SYN 13-OCT-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6176)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 1, 2017 from SnapGene 4.0.5
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6176
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    114..133
                     /label=Verification forward (VF2) primer binding site
                     /note="/ApEinfo_fwdcolor=#85dae9
                     /ApEinfo_revcolor=#85dae9"
     terminator      184..227
                     /label=bacterial terminator
                     /note="/ApEinfo_fwdcolor=#c6c9d1
                     /ApEinfo_revcolor=#c6c9d1"
     misc_feature    230..249
                     /label=BioBrick prefix CDS
                     /label=BioBrick prefix
                     /note="/Biobrick prefix for coding sequences
                     /ApEinfo_fwdcolor=#84b0dc
                     /ApEinfo_revcolor=#84b0dc"
     CDS             251..4357
                     /codon_start=1
                     /label=dCas9VRER
                     /note="/ApEinfo_fwdcolor=#f58a5e
                     /ApEinfo_revcolor=#f58a5e"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKEYRSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     misc_feature    278..280
                     /label=D10A
                     /label=dCas9
                     /note="/ApEinfo_fwdcolor=#ff9ccd
                     /ApEinfo_revcolor=#ff9ccd"
     misc_feature    1594
                     /label=T->A
                     /note="/ApEinfo_fwdcolor=#f58a5e
                     /ApEinfo_revcolor=#f58a5e"
     misc_feature    2768..2770
                     /label=H840A
                     /label=dCas9
                     /note="/ApEinfo_fwdcolor=#ff9ccd
                     /ApEinfo_revcolor=#ff9ccd"
     misc_feature    3629..4377
                     /label=gBlock_dCas9
     misc_feature    3653..3655
                     /label=D1335V
                     /note="/ApEinfo_fwdcolor=#85dae9
                     /ApEinfo_revcolor=#85dae9"
     misc_feature    3902..3904
                     /label=G1218R
                     /note="/ApEinfo_fwdcolor=#c6c9d1
                     /ApEinfo_revcolor=#c6c9d1"
     misc_feature    4253..4255
                     /label=R1335E
                     /note="/ApEinfo_fwdcolor=#ff9ccd
                     /ApEinfo_revcolor=#ff9ccd"
     misc_feature    4259..4261
                     /label=T1337R
                     /note="/ApEinfo_fwdcolor=#c7b0e3
                     /ApEinfo_revcolor=#c7b0e3"
     misc_feature    4358..4378
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     stem_loop       4379..4450
                     /label=E. coli his operon terminator
                     /note="/ApEinfo_fwdcolor=#b7e6d7
                     /ApEinfo_revcolor=#b7e6d7"
     misc_feature    complement(4514..4533)
                     /label=Verification reverse (VR) primer binding site
                     /note="/ApEinfo_fwdcolor=#84b0dc
                     /ApEinfo_revcolor=#84b0dc"
     misc_feature    complement(4618..5232)
                     /label=rep (pMB1)
                     /note="/ApEinfo_fwdcolor=#f8d3a9
                     /ApEinfo_revcolor=#f8d3a9"
     terminator      5402..5496
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5517..6176)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
       61 tatttcatta tggtgaaagt tggaacctct tacgtgcccg atcaactcga gtgccacctg
      121 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
      181 agaatttcag ataaaaaaaa tccttagctt tcgctaagga tgatttctgg aattcgcggc
      241 cgcttctaga atggataaga aatactcaat aggcttagct atcggcacaa atagcgtcgg
      301 atgggcggtg atcactgatg aatataaggt tccgtctaaa aagttcaagg ttctgggaaa
      361 tacagaccgc cacagtatca aaaaaaatct tataggggct cttttatttg acagtggaga
      421 gacagcggaa gcgactcgtc tcaaacggac agctcgtaga aggtatacac gtcggaagaa
      481 tcgtatttgt tatctacagg agattttttc aaatgagatg gcgaaagtag atgatagttt
      541 ctttcatcga cttgaagagt cttttttggt ggaagaagac aagaagcatg aacgtcatcc
      601 tatttttgga aatatagtag atgaagttgc ttatcatgag aaatatccaa ctatctatca
      661 tctgcgaaaa aaattggtag attctactga taaagcggat ttgcgcttaa tctatttggc
      721 cttagcgcat atgattaagt ttcgtggtca ttttttgatt gagggagatt taaatcctga
      781 taatagtgat gtggacaaac tatttatcca gttggtacaa acctacaatc aattatttga
      841 agaaaaccct attaacgcaa gtggagtaga tgctaaagcg attctttctg cacgattgag
      901 taaatcaaga cgattagaaa atctcattgc tcagctcccc ggtgagaaga aaaatggctt
      961 atttgggaat ctcattgctt tgtcattggg tttgacccct aattttaaat caaattttga
     1021 tttggcagaa gatgctaaat tacagctttc aaaagatact tacgatgatg atttagataa
     1081 tttattggcg caaattggag atcaatatgc tgatttgttt ttggcagcta agaatttatc
     1141 agatgctatt ttactttcag atatcctaag agtaaatact gaaataacta aggctcccct
     1201 atcagcttca atgattaaac gctacgatga acatcatcaa gacttgactc ttttaaaagc
     1261 tttagttcga caacaacttc cagaaaagta taaagaaatc ttttttgatc aatcaaaaaa
     1321 cggatatgca ggttatattg atgggggagc tagccaagaa gaattttata aatttatcaa
     1381 accaatttta gaaaaaatgg atggtactga ggaattattg gtgaaactaa atcgtgaaga
     1441 tttgctgcgc aagcaacgga cctttgacaa cggctctatt ccccatcaaa ttcacttggg
     1501 tgagctgcat gctattttga gaagacaaga agacttttat ccatttttaa aagacaatcg
     1561 tgagaagatt gaaaaaatct tgacttttcg aataccttat tatgttggtc cattggcgcg
     1621 tggcaatagt cgttttgcat ggatgactcg gaagtctgaa gaaacaatta ccccatggaa
     1681 ttttgaagaa gttgtcgata aaggtgcttc agctcaatca tttattgaac gcatgacaaa
     1741 ctttgataaa aatcttccaa atgaaaaagt actaccaaaa catagtttgc tttatgagta
     1801 ttttacggtt tataacgaat tgacaaaggt caaatatgtt actgaaggaa tgcgaaaacc
     1861 agcatttctt tcaggtgaac agaagaaagc cattgttgat ttactcttca aaacaaatcg
     1921 aaaagtaacc gttaagcaat taaaagaaga ttatttcaaa aaaatagaat gttttgatag
     1981 tgttgaaatt tcaggagttg aagatagatt taatgcttca ttaggtacct accatgattt
     2041 gctaaaaatt attaaagata aagatttttt ggataatgaa gaaaatgaag atatcttaga
     2101 ggatattgtt ttaacattga ccttatttga agatagggag atgattgagg aaagacttaa
     2161 aacatatgct cacctctttg atgataaggt gatgaaacag cttaaacgtc gccgttatac
     2221 tggttgggga cgtttgtctc gaaaattgat taatggtatt agggataagc aatctggcaa
     2281 aacaatatta gattttttga aatcagatgg ttttgccaat cgcaatttta tgcagctgat
     2341 ccatgatgat agtttgacat ttaaagaaga cattcaaaaa gcacaagtgt ctggacaagg
     2401 cgatagttta catgaacata ttgcaaattt agctggtagc cctgctatta aaaaaggtat
     2461 tttacagact gtaaaagttg ttgatgaatt ggtcaaagta atggggcggc ataagccaga
     2521 aaatatcgtt attgaaatgg cacgtgaaaa tcagacaact caaaagggcc agaaaaattc
     2581 gcgagagcgt atgaaacgaa tcgaagaagg tatcaaagaa ttaggaagtc agattcttaa
     2641 agagcatcct gttgaaaata ctcaattgca aaatgaaaag ctctatctct attatctcca
     2701 aaatggaaga gacatgtatg tggaccaaga attagatatt aatcgtttaa gtgattatga
     2761 tgtcgatgcc attgttccac aaagtttcct taaagacgat tcaatagaca ataaggtctt
     2821 aacgcgttct gataaaaatc gtggtaaatc ggataacgtt ccaagtgaag aagtagtcaa
     2881 aaagatgaaa aactattgga gacaacttct aaacgccaag ttaatcactc aacgtaagtt
     2941 tgataattta acgaaagctg aacgtggagg tttgagtgaa cttgataaag ctggttttat
     3001 caaacgccaa ttggttgaaa ctcgccaaat cactaagcat gtggcacaaa ttttggatag
     3061 tcgcatgaat actaaatacg atgaaaatga taaacttatt cgagaggtta aagtgattac
     3121 cttaaaatct aaattagttt ctgacttccg aaaagatttc caattctata aagtacgtga
     3181 gattaacaat taccatcatg cccatgatgc gtatctaaat gccgtcgttg gaactgcttt
     3241 gattaagaaa tatccaaaac ttgaatcgga gtttgtctat ggtgattata aagtttatga
     3301 tgttcgtaaa atgattgcta agtctgagca agaaataggc aaagcaaccg caaaatattt
     3361 cttttactct aatatcatga acttcttcaa aacagaaatt acacttgcaa atggagagat
     3421 tcgcaaacgc cctctaatcg aaactaatgg ggaaactgga gaaattgtct gggataaagg
     3481 gcgagatttt gccacagtgc gcaaagtatt gtccatgccc caagtcaata ttgtcaagaa
     3541 aacagaagta cagacaggcg gattctccaa ggagtcaatt ttaccaaaaa gaaattcgga
     3601 caagcttatt gctcgtaaaa aagactggga tccaaaaaaa tatggtggtt ttgttagtcc
     3661 aacggtagct tattcagtcc tagtggttgc taaggtggaa aaagggaaat cgaagaagtt
     3721 aaaatccgtt aaagagttac tagggatcac aattatggaa agaagttcct ttgaaaaaaa
     3781 tccgattgac tttttagaag ctaaaggata taaggaagtt aaaaaagact taatcattaa
     3841 actacctaaa tatagtcttt ttgagttaga aaacggtcgt aaacggatgc tggctagtgc
     3901 ccgagaatta caaaaaggaa atgagctggc tctgccaagc aaatatgtga attttttata
     3961 tttagctagt cattatgaaa agttgaaggg tagtccagaa gataacgaac aaaaacaatt
     4021 gtttgtggag cagcataagc attatttaga tgagattatt gagcaaatca gtgaattttc
     4081 taagcgtgtt attttagcag atgccaattt agataaagtt cttagtgcat ataacaaaca
     4141 tagagacaaa ccaatacgtg aacaagcaga aaatattatt catttattta cgttgacgaa
     4201 tcttggagct cccgctgctt ttaaatattt tgatacaaca attgatcgta aagaatatcg
     4261 ttctacaaaa gaagttttag atgccactct tatccatcaa tccatcactg gtctttatga
     4321 aacacgcatt gatttgagtc agctaggagg tgactgatac tagtagcggc cgctgcagtc
     4381 cggcaaaaaa gggcaaggtg tcaccaccct gccctttttc tttaaaaccg aaaagattac
     4441 ttcgcgttat gcaggcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4501 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4561 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4621 tgctggcgtt tttccacagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4681 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4741 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4801 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4861 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4921 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4981 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5041 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5101 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5161 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5221 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5281 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5341 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagc tcgaggcttg
     5401 gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga
     5461 gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgat atcaaattac
     5521 gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg
     5581 aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct
     5641 tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg
     5701 tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca
     5761 ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat
     5821 atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca
     5881 gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg
     5941 tctttcattg ccatacgaaa ttccggatga gcattcatca ggcgggcaag aatgtgaata
     6001 aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc
     6061 agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct
     6121 ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt ctccat
//
